W/F Purple team Note: For our unequal ratio test, we forgot to add the diluted lipofectamine to this sample when the rest of the samples received lipofectamine. We added the 50 ul and let it incubate for as long as possible, but that was only about 30 seconds before we had to finish the experiment and clean up to get to our next section.

W/F Purple team Note: For our unequal ratio test, we forgot to add the diluted lipofectamine to this sample when the rest of the samples received lipofectamine. We added the 50 ul and let it incubate for as long as possible, but that was only about 30 seconds before we had to finish the experiment and clean up to get to our next section.

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====W/F====

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Current revision

Contents

Schedule

Sign up for a time to do flow cytometry in the table below. Please arrive at the time you choose and then expect to spend about 45 minutes in TC and another 45 minutes at FACS. TWO or THREE teams should sign up for each time slot.

T/R

Time block

Team 1

Team 2

Team 3

I: 1-2:30 pm

Blue

Platinum

White

II: 2-3:30-pm

Purple

Yellow

Green

III: 3-4:30 pm

Orange

Pink

Red

W/F

Time block

Team 1

Team 2

(Team 3)

I: 1-2:30 pm

Red Team

Orange Team

White Team

II: 2-3:30-pm

Pink Team

Blue Team

III: 3-4:30 pm

Purple Team

Green Team

Data Summary

Key general notes about experiment

Difference between T/R and W/F samples

On T/R, plain media wells were exchanged to fresh media, while wells with DMSO or C401 just stayed in the same media that had been added to the plate at 10am on the morning of M1D6.

On W/F, all three types of media were available and exchanged to fresh before transfection.

Collection time consideration

As discussed with most groups, we collected at a high flowrate in order to test many samples.

The downside is that transient fluctuations in the data can have a big influence, and may not be averaged out.

Keep this trade-off in mind as you look at personal duplicates as well as class-wide data.

Reported data

To aid class-wide comparisons (and statistics!), please post the requested information in the table below in a timely fashion using three significant digits (ex. 3.72). For samples done in duplicate, please separate the two values with a semi-colon. Once you've completed your statistics, don't get over-excited with the number of significant figures you report! For example, "3.7" will suffice. Please describe the ratio test condition in the form "0.2 ug D5 and 0.05 ug CUT D3."

T/R

Team color

% R2 gated mock

" delta 5 (D5)

" delta 3 (D3)

" EGFP

" EGFP + DMSO

" D5 + D3

" D5 + cut D3

" D5 + cut D3 + DMSO

" D5 + cut D3 + C401

% R2 Ratio test

Ratio condition

Purple

0.00

0.00

0.00

67.0; 68.6

60.9; 61.3

3.91; 3.39

11.3; 13.8

9.94; 8.90

6.86; 7.34

11.2; 14.9

1:2; 0.1ug D5 and 0.2ug CUT D3

Platinum

0.01

0.00

0.01

68.9; 68.8

69.6; 70.5

4.02; 3.72

11.2; 9.98

8.56; 7.57

4.24; 5.45

14.74; 13.93

1:3; 0.1ug D5 and 0.3ug cut D3

Yellow

0.00

0.00

0.00

71.61; 73.35

76.07; 76.07

4.06; 2.93

13.30; 11.93

11.28; 9.26

2.46; Lost Sample

15.00; 12.78

1:3; 0.1µg ∆5 and 0.3µg cut ∆3

White

0.00

0.00

0.00

76.57; 77.87

76.82; 77.56

3.14; 2.71

7.36; 7.19

6.32; 5.21

3.15; 3.36

8.24; 7.86

1:3; 0.05µg ∆5 and 0.15µg cut ∆3

Pink

0.00

0.02

0.00

55.70; 55.02

64.04; 65.66

3.47; 2.41

5.22; 4.72

3.59; 2.78

1.43; 1.81

2.64; 1.24

4:1, 0.2µg ∆5 and 0.05µg digested ∆3

Orange

0.00

0.02

0.00

71.06; 75.39

69.00; 74.31

2.87; 2.03

11.41; 11.93

9.77; 9.56

7.49; 7.09

17.03; 15.42

4:1, 0.4 µg D5 and 0.1 µg CUT D3

Green

0.00

0.00

0.00

71.12,71.19

64.79,70.18

2.51,2.26

8.98,8.48

7.66,5.86

3.59,2.63

12.10,12.02

1:5, 0.1µg ∆5 and 0.5µg digested ∆3

Blue

0.00

0.00

0.00

48.0, 53.4

51.11, 57.83

2.02, 1.87

10.63, 8.88

7.89, 7.80

3.88, 4.16

1.83, 1.71

1:4 0.1µg ∆5 and 0.4µg digested ∆3

Red

0.00

0.00

0.00

65.17, 66.74

60.46, 63.72

2.47, 2.80

7.50, 8.51

6.85, 6.36

2.74, 3.72

14.00, 16.63

4:1 0.4µg ∆5 and 0.1µg digested ∆3

If you did not collect the default amount of R1 cells (10,000) per sample, please list your cell count below:

Pink Team Observations: There was something wrong with our digested ∆3-plasmid, because we have a smaller percentage of fluoroscence compared to other teams.

W/F Purple team Note: For our unequal ratio test, we forgot to add the diluted lipofectamine to this sample when the rest of the samples received lipofectamine. We added the 50 ul and let it incubate for as long as possible, but that was only about 30 seconds before we had to finish the experiment and clean up to get to our next section.
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W/F

Team color

% R2 gated mock

" delta 5 (D5)

" delta 3 (D3)

" EGFP

" EGFP + DMSO

" D5 + D3

" D5 + cut D3

" D5 + cut D3 + DMSO

" D5 + cut D3 + C401

% R2 Ratio test

Ratio condition

Yellow

0.0

0.0

0.0

66.0; 63.49

53.66; 57.27

0.51; 0.47

12.04; 10.01

5.26; 10.37

8.02; 6.54

0.67; 0.76

10:1 ; 0.02 ug Delta 5, 0.2 ug Cut-Delta 3

Pink

0.0

0.0

0.0

71.3; 66.4

71.9; 72.8

2.5; 2.8

6.4; 6.2

5.1; 5.5

3.5; 2.8

7.2; 5.8

10:1 ; 5ug cut-del3, 0.5ug del5

Red

0.0

0.0

0.0

0.0; 0.0 (Our GFP solution didn't include carrier)

0.0; 0.0

2.88; 2.28

12.48; 10.32

12.24; 11.26

6.66; 5.48

6.92; 8.43

1:5 0.1ug D5 and 0.5ug cut D3

Purple

0.0

0.01

0.0

57.7; 61.5

62.3; 52.7

2.42; 1.78

8.54; 7.81

9.01; 7.09

5.35; 6.33

4.30; 1.33

1:5 .025 ug of Δ5 and .125 ug of digested Δ3

Green

0.0

0.0

0.0

51.54; 56.23

66.78; 59.49

2.46; 2.48

5.33; 4.30

6.10; 4.51

3.77; 4.40

2.44; 18.28

10:1; .5 ug D5: .05 ug digested D3

Blue

0.0

0.0

0.0

59.7; 68.0

63.5; 58.9

1.9; 1.5; 2.4

9.4; 9.4

12.0

7.0; 5.9

14.5; 15.0

5:1; 0.5 µg D5: 0.1 µg digested D3

Orange

0.0

0.0

0.0

66.9; 63.4

70.2; 68.5

2.85; 2.2

9.2; 7.2

11.8; 11.01

6.84; 6.47

14.2; 12.54

10:1 .1 ug D5: .01 ug of D3

White

0.0

0.0

0.0

61.21; 62.41

60.17; 61.49

2.53; 2.56

8.53; 8.43

9.17; 7.07

5.71; 6.73

10.99; 10.66

1:2; 0.1 ug D5: 0.2 ug digested D3

If you did not collect the default amount of R1 cells (10,000) per sample, please list your cell count below:

Blue: Collected 3 samples of D5+D3, because we had leftover D5+D3 solution so we plated an extra well of it. Only collected one sample of D5+cut D3+DMSO because we forgot to add more media when we were changing the media before adding the appropriate samples, so the cells died.